| Product Includes | Product # | Volume | Cap Color | Storage Temp |
|---|---|---|---|---|
| Phospho-4E-BP1 (Thr37/46) Capture Rabbit mAb (100X) | 18033 | 400 µl |
|
+4C |
| 4E-BP1 Detection Mouse mAb (100X) | 40141 | 400 µl |
|
+4C |
| Anti-mouse IgG, HRP-linked Antibody (1000X) | 16736 | 40 µl |
|
-20C |
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Description
CST's PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Kit #7216. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Phospho-4E-BP1 (Thr37/Thr46) Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a 4E-BP1 Detection Antibody and anti-Mouse IgG, HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of Phospho-4E-BP1 (Thr37/Thr46) protein.
Antibodies in kit are custom formulations specific to kit.
Reagents not supplied
Phosphate Buffered Saline with Tween-20 (PBST-20X) #9809
Cell Lysis Buffer (10X) #9803
TMB Substrate #7004
STOP Solution #7002
Blocking Buffer: 1X PBS/0.5% Tween-20, 1% BSA
96 Well Microplates**
Microplate Reader
** Antibody Pairs have been validated on Corning© 96 Well Clear Polystyrene High Bind Stripwell™ Microplates (#2592).
Notes: Antibody pairs have been optimized using recommended buffers, reagents, plates and the included protocol. Solutions should be made fresh daily.
Background
Translation repressor protein 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation by binding to the translation initiation factor eIF4E. Hyperphosphorylation of 4E-BP1 disrupts this interaction and results in activation of cap-dependent translation (1). Both the PI3 kinase/Akt pathway and FRAP/mTOR kinase regulate 4E-BP1 activity (2,3). Multiple 4E-BP1 residues are phosphorylated in vivo (4). While phosphorylation by FRAP/mTOR at Thr37 and Thr46 does not prevent the binding of 4E-BP1 to eIF4E, it is thought to prime 4E-BP1 for subsequent phosphorylation at Ser65 and Thr70 (5).
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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